安宫牛黄丸干预对创伤性颅脑损伤大鼠学习记忆能力的影响

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目的:探讨安宫牛黄丸干预对创伤性颅脑损伤大鼠学习记忆能力的影响。方法:采用随机数字表将40只健康清洁级SD雄性大鼠随机分为假手术组(假手术+1 ml蒸馏水)、创伤组(造模+1 ml蒸馏水)、安宫牛黄丸低浓度组(造模+安宫牛黄丸2 g/kg)、安宫牛黄丸高浓度组(造模+安宫牛黄丸3 g/kg)。避暗实验检测避暗潜伏期和错误次数;取大鼠脑组织,黄嘌呤酶酶法测定超氧化物歧化酶(SOD)活性,硫代巴比妥酸法检测丙二醛(MDA)含量,苏木精-伊红(HE)染色观察细胞结构改变,蛋白免疫印迹法(Western blot)检测tau蛋白及其磷酸化蛋白表达水平。实验数据经正态性检验后进行方差分析和n t检验。n 结果:创伤后第18小时,安宫牛黄丸低浓度组、安宫牛黄丸高浓度组大鼠避暗潜伏期分别为(227.65±21.46)、(250.37±26.05) s,明显长于创伤组[(174.09±34.13) s],错误次数分别为(1.95±0.63)、(1.17±0.49)次,明显少于创伤组[(3.80±0.77)次,n t=22.039、24.726,12.404、14.837,n P<0.05],差异有统计学意义,安宫牛黄丸高浓度组改善效果更明显(n t=11.827、6.720,n P<0.05)。安宫牛黄丸低浓度组、安宫牛黄丸高浓度组大鼠脑组织中SOD活性分别为(270.25±21.60)、(314.97±23.14) U/mg蛋白,明显高于创伤组[(128.71±19.23) U/mg蛋白],MDA含量分别为(8.51±0.94)、(6.05±0.61) μmol/mg蛋白,明显低于创伤组[(11.74±1.20) μmol/mg蛋白,n t=20.731、24.209,7.527、8.398,n P<0.05],差异有统计学意义,安宫牛黄丸高浓度组改善效果更明显(n t=12.694、5.352,n P<0.05),差异有统计学意义。大鼠脑脑组织中tau、p-tau S199、p-tau S404蛋白表达在安宫牛黄丸低浓度组中分别为1.73±0.29、0.93±0.11、1.42±0.25,安宫牛黄丸高浓度组中分别为1.05±0.17、0.67±0.08、0.95±0.18,均明显低于创伤组(n t=8.128、9.631、9.305,13.520、12.758、11.207,n P<0.05),差异有统计学意义,安宫牛黄丸高浓度组降低幅度更明显(n t=7.392、5.813、6.605,n P<0.05),差异有统计学意义。n 结论:安宫牛黄丸可明显改善创伤性颅脑损伤大鼠的学习记忆能力,可能与抑制脑组织氧自由基反应和tau蛋白磷酸化水平有关。“,”Objective:To explore the effect of Angong Niuhuang pill on the learning and memory ability of rats with traumatic brain injury.Methods:Using a random number table, a total of 40 healthy and clean SD male rats were randomly divided into four groups, sham operation group (sham operation+ 1 ml distilled water), trauma group (modeling+ 1 ml distilled water), Angong Niuhuang pill low concentration group (modeling+ Angong Niuhuang pill 2 g/kg), Angong Niuhuang pill high concentration group (modeling+ Angong Niuhuang pill 3 g/kg). The escape latency and the number of errors were detected by the escape darkness experiment. The brain tissues of rats were taken, the activity of superoxide dismutase (SOD) was measured by xanthinase method, and the content of malondialdehyde (MDA) was measured by thiobarbituric acid method. Hematoxylin and eosin (HE) staining was used to observe the changes of cell structure. The expression of tau protein and its phosphorylation protein were detected by Western blotting. The data were analyzed by ANOVA and n t test.n Results:On the 18th day after trauma, the escape latency of rats in Angong Niuhuang pill low concentration group and Angong Niuhuang pill high concentration group was (227.65±21.46), (250.37±26.05) s respectively, which was significantly longer than that in trauma group [(174.09±34.13) s], and the number of errors were (1.95±0.63), (1.17±0.49) times respectively, which were significantly less than those in trauma group [(3.80±0.77) times, n t=22.039, 24.726, 12.404, 14.837, n P<0.05]. The improvement effect of Angong Niuhuang pill high concentration group was more obvious (n t=11.827, 6.720, n P<0.05). The SOD activity in the brain tissue of rats in the Angong Niuhuang pill low concentration group and the Angong Niuhuang pill high concentration group was (270.25±21.60), (314.97±23.14) U/mgpro respectively, which was significantly higher than in the trauma group [(128.71±19.23) U/mgpro] and MDA content was (8.51±0.94), (6.05±0.61) μmol/mgpro, which was significantly lower than in the trauma group [(11.74±1.20) μmol/mgpro,n t=20.731, 24.209, 7.527, 8.398, n P<0.05]. The improvement effect of Angong Niuhuang pill high concentration group was more obvious (n t=12.694, 5.352, n P<0.05). The expression levels of tau, p-tau S199, and p-tau S404 in the brain tissue of rats were 1.73±0.29, 0.93±0.11, 1.42±0.25 in the Angong Niuhuang pill low-concentration group, respectively, and in the Angong Niuhuang pill high-concentration group they were 1.05±0.17, 0.67±0.08, 0.95±0.18 respectively, which were significantly lower than those in the trauma group (n t=8.128, 9.631, 9.305, 13.520, 12.758, 11.207, n P<0.05). The improvement effect of Angong Niuhuang pill high concentration group was more obvious (n t=7.392, 5.813, 6.605, n P<0.05).n Conclusion:Angong Niuhuang pill can significantly improve the learning and memory ability of rats with traumatic brain injury, which may be related to the inhibition of oxygen free radical reaction and tau protein phosphorylation level.
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